Effects of metalloporphyrin catalytic antioxidants in experimental brain ischemia

Reactive oxygen species play a role in the response of brain to ischemia. The effects of metalloporphyrin catalytic antioxidants (AEOL 10113 and AEOL 10150) were examined after murine middle cerebral artery occlusion (MCAO). Ninety minutes after reperfusion from 90 min MCAO in the rat, AEOL 10113, AEOL 10150, or vehicle were given intracerebroventricularly. AEOL 10113 and AEOL 10150 similarly reduced infarct size (35%) and neurologic deficit. AEOL 10113 caused behavioral side effects at twice the neuroprotective dose while AEOL 10150 required a 15-fold increase from the neuroprotective dose to cause behavioral changes. AEOL 10150, given 6 h after 90 min MCAO, reduced total infarct size by 43% without temperature effects. Brain AEOL 10150 elimination t(1/2) was 10 h. In the mouse, intravenous AEOL 10150 infusion post-MCAO reduced both infarct size (25%) and neurologic deficit. Brain AEOL 10150 uptake, greater in the ischemic hemisphere, was dose- and time-dependent. AEOL 10150 had direct effects on proteomic events and ameliorated changes caused by ischemia. In primary mixed neuronal/glial cultures exposed to 2 h of O(2)/glucose deprivation, AEOL 10150 reduced lactate dehydrogenase release dose-dependently and selectively preserved aconitase activity in concentrations consistent with neuroprotection in vivo. AEOL 10150 is an effective neuroprotective compound offering a wide therapeutic window with a large margin of safety against adverse behavioral side effects.     

Acetylcholine and Dopamine Promote the Production of Platelet Activating Factor in Immature Cells of Chick Embryonic Retina

We have previously shown that platelet-activating factor (PAF), a naturally occurring lipid mediator of cell-to-cell communication, was produced by 3-day-old chick retina stimulated with acetylcholine (ACh) and dopamine (DA), but not with other neurotransmitters. ACh and DA stimulated PAF synthesis via a dithiothreitol (DTT)-insensitive cholinephosphotransferase, without affecting the acetyltransferase pathway, which was stimulated only by the calcium ionophore A23187. Therefore, we attempted to study the effects of neurotransmitters on PAF production and on the activities of the DTT-insensitive cholinephosphotransferase and acetyltransferase in the developing chick embryo retina up to hatching. Our results show that PAF was produced already at 8 days of development, when retinal cells are still rather immature and ganglion and Mueller cells are the only differentiated cells. The stimulation of PAF production occurred with ACh and not with other neurotransmitters. In older stages, DA also stimulated PAF production, as already described in the chick after hatching. DTT-insensitive cholinephosphotransferase and acetyltransferase activities were present in 8-day-old embryos, the earliest stage analyzed. Both enzymatic activities increased with age; DTT-insensitive cholinephosphotransferase increased rapidly from day 12 up to day 18, whereas acetyltransferase activity increased linearly up to the time of hatching. To promote PAF production, ACh and DA activate DTT-insensitive cholinephosphotransferase, but not acetyltransferase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Erratum to: Genomics-Guided Precise Anti-Epileptic Drug Development

Neurochemical Research -     

Species limits and morphometric and environmental variation within the South Andean and Patagonian Mulinum spinosum species-group (Apiaceae-Azorelloideae)

Delimiting species is an important, but frequently difficult aspect of systematics that should be addressed using data from multiple sources. Here we combine morphometric analyses and environmental characteristics to delimit species in the South Andean and Patagonian taxonomically difficult species-group composed by Mulinum spinosum, M. echinus and M. leptacanthum (Apiaceae-Azorelloideae). Molecular phylogenies have shown that these three species form part of a polytomy together with other Mulinum species, and therefore these data are not useful for their delimitation. We include measurements of 25 morphometric variables from 163 herbarium specimens and perform univariate and multivariate principal component analysis (PCA) and discriminant analysis (DA) to establish the limits amongst the three mostly sympatric, morphologically similar, and phylogenetically unresolved species. We also use 19 bioclimatic and three geographic variables from localities of the specimens to infer environmental characteristics of the taxa and test their relation with morphological variation. Morphological evidence supports the inclusion of M. echinus within the morphologically and climatically variable M. spinosum, and rejects its recognition as a distinct taxon at any rank. On the contrary, M. leptacanthum is considered a morphologically distinct species, generally restricted to high altitude areas of the southern Andes with a cooler and wetter climate. Within the widespread M. spinosum, environmental gradients of precipitation and temperature relate to morphological gradients (e.g., in leaf and inflorescence sizes, in leaf acicularity, and in fruit-wing width). These last results showed that the large morphological variation in vegetative and reproductive characteristics of this species that grows in arid and semiarid habitats are related to regional climatic gradients that have probably been important in the evolution of this species' plasticity, diversification, and differentiation.     

Suppression of late-flowering and semi-dwarf phenotypes in the arabidopsis clock mutant lhy-12;cca1-101 by phyB under continuous light

Photoperiodic flowering in Arabidopsis is controlled not only by floral activators such as GI, CO and FT, but also by repressors such as SVP and FLC. Double mutations in LHY and CCA1 (lhy;cca1) accelerated flowering under short days, mainly by the GI-CO dependent pathway. In contrast, lhy;cca1 showed delayed flowering under continuous light (LL), probably due to the GI-CO independent pathway. This late-flowering phenotype was suppressed by svp, flc and elf3. However, how SVP, FLC and ELF3 mediate LHY/CCA1 and flowering time is not fully understood. We found that lhy;cca1 exhibited short hypocotyls and petioles under LL, but the molecular mechanism for these effects has not been elucidated.To address these questions, we performed a screen for mutations that suppress either or both of the lhy;cca1 phenotypes under LL, using two different approaches. We identified two novel mutations, a dominant (del1) and a recessive (phyB-2511) allele of phyB. The flowering times of single mutants of three phyB alleles, hy3-1, del1 and phyB-2511, are almost the same and earlier than those of wild-type plants. A similar level of acceleration of flowering time was observed in all three phyB mutants tested when combined with the late-flowering mutations co-2 and SVPox. However, the effect of phyB-2511 on lhy;cca1 was different from those by hy3-1 or del1. svp-3 did not strongly enhance the early-flowering phenotypes of phyB-2511 or del1. These results suggest that light signaling via PhyB may affect factors downstream of the clock proteins, controlling flowering time and organ elongation. phyB mutations with different levels of effects on lhy;cca1-dependent late flowering would be useful to determine a specific role for PHYB in the flowering pathway controlled by lhy;cca1 under LL.Key words: Arabidopsis thaliana, CCA1, circadian clock, CO, FT, LHY, organ elongation, photoperiodic flowering, PHYB, SVP     

MiR-34a/c-Dependent PDGFR-α/β Downregulation Inhibits Tumorigenesis and Enhances TRAIL-Induced Apoptosis in Lung Cancer

Lung cancer is the leading cause of cancer mortality in the world today. Although some advances in lung cancer therapy have been made, patient survival is still poor. MicroRNAs (miRNAs) can act as oncogenes or tumor-suppressor genes in human malignancy. The miR-34 family consists of tumor-suppressive miRNAs, and its reduced expression has been reported in various cancers, including non-small cell lung cancer (NSCLC). In this study, we found that miR-34a and miR-34c target platelet-derived growth factor receptor alpha and beta (PDGFR-α and PDGFR-β), cell surface tyrosine kinase receptors that induce proliferation, migration and invasion in cancer. MiR-34a and miR-34c were downregulated in lung tumors compared to normal tissues. Moreover, we identified an inverse correlation between PDGFR-α/β and miR-34a/c expression in lung tumor samples. Finally, miR-34a/c overexpression or downregulation of PDGFR-α/β by siRNAs, strongly augmented the response to TNF-related apoptosis inducing ligand (TRAIL) while reducing migratory and invasive capacity of NSCLC cells.     

Mechanical properties of the triceps surae tendon and aponeurosis in relation to intensity of sport activity

The purpose of the present study was to investigate whether the mechanical properties (i.e. force strain relationship) of the triceps surae tendon and aponeurosis relate to the performed sport activity in an intensity-dependent manner. This was done by comparing sprinters with endurance runners and subjects not active in sports. Sixty-six young male subjects (26+/-5 yr; 183+/-6 cm; 77.6+/-6.7 kg) participated in the study. Ten of these subjects were adults not active in sports, 28 were endurance runners and 28 sprinters. All subjects performed isometric maximal voluntary plantar flexion contractions (MVC) on a dynamometer. The distal aponeuroses of the gastrocnemius medialis (GM) was visualised by ultrasound during the MVC. The results showed that only the sprinters had higher normalised stiffness (relationship between tendon force and tendon strain) of the triceps surae tendon and aponeurosis and maximal calculated tendon forces than the endurance runners and the subjects not active in sports. Furthermore, including the data of all 66 examined participants tendon stiffness correlated significantly (r=0.817, P<0.001) with the maximal tendon force achieved during the MVC. It has been concluded that the mechanical properties of the triceps surae tendon and aponeurosis do not show a graded response to the intensity of the performed sport activity but rather remain at control level in a wide range of applied strains and that strain amplitude and/or frequency should exceed a given threshold in order to trigger additional adaptation effects. The results further indicate that subjects with higher muscle strength possibly increase the margin of tolerated mechanical loading of the tendon due to the greater stiffness of their triceps surae tendon and aponeurosis.